Wang Dingxue, Huang Wenqi, Li Gao
Oncology department, the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang, China.
Biomol Biomed. 2025 Apr 3;25(5):1184-1196. doi: 10.17305/bb.2024.11209.
Hepatocellular carcinoma (HCC) exhibits a subtle onset, high incidence rates, and low survival rates, becoming a substantial threat to human health. Hence, it is crucial to discover fresh biomarkers and treatment targets for the early detection and management of HCC. CCK-8, scratch-wound, and transwell assays were used to evaluate the biological properties of HCC cell lines (Huh-7 and Hep3B). Bioinformatics analysis identified the downstream target mRNA of miR-145-5p as acyl-CoA synthetase long-chain family member 4 (ACSL4). RT-qPCR was used to test miR-145-5p and ACSL4 levels. Transwell chambers were used to co-incubate purified CD8+ T cells and HCC cells for 48 h, and the effect of CD8+ T cells on apoptosis in HCC cells was detected by flow cytometry. A subcutaneous graft tumor model was constructed, and ELISA kits were used to assess cytokine levels and CD8+ T cell activation markers. HCC cells showed a decline in miR-145-5p levels and a rise in ACSL4 levels. Overexpression of miR-145-5p hindered HCC cell proliferation, migration, and invasion, while stimulating CD8+ T cell activation. Conversely, overexpression of ACSL4 enhanced the malignant biological properties of HCC cells and reduced the effect of CD8+ T cells, while silencing ACSL4 had the opposite effect. miR-145-5p targeted and downregulated ACSL4, while overexpression of miR-145-5p weakened the promotion of HCC malignant progression caused by ACSL4 overexpression. Additionally, overexpression of miR-145-5p and silencing ACSL4 were effective in inhibiting tumor growth in vivo. In conclusion, miR-145-5p targets and downregulates ACSL4, leading to the inhibition of HCC malignant progression and preventing immune escape in HCC cells.
肝细胞癌（HCC）起病隐匿，发病率高，生存率低，对人类健康构成重大威胁。因此，发现新的生物标志物和治疗靶点对于HCC的早期检测和管理至关重要。采用CCK-8、划痕试验和Transwell试验评估肝癌细胞系（Huh-7和Hep3B）的生物学特性。生物信息学分析确定miR-145-5p的下游靶mRNA为酰基辅酶A合成酶长链家族成员4（ACSL4）。采用RT-qPCR检测miR-145-5p和ACSL4水平。使用Transwell小室将纯化的CD8+T细胞与HCC细胞共孵育48小时，通过流式细胞术检测CD8+T细胞对HCC细胞凋亡的影响。构建皮下移植瘤模型，使用ELISA试剂盒评估细胞因子水平和CD8+T细胞活化标志物。HCC细胞中miR-145-5p水平下降，ACSL4水平上升。miR-145-5p的过表达抑制了HCC细胞的增殖、迁移和侵袭，同时刺激了CD8+T细胞的活化。相反，ACSL4的过表达增强了HCC细胞的恶性生物学特性，降低了CD8+T细胞的作用，而沉默ACSL4则产生相反的效果。miR-145-5p靶向并下调ACSL4，而miR-145-5p的过表达减弱了ACSL4过表达对HCC恶性进展的促进作用。此外，miR-145-5p的过表达和ACSL4的沉默在体内有效抑制肿瘤生长。总之，miR-145-5p靶向并下调ACSL4，导致HCC恶性进展受到抑制，并防止HCC细胞发生免疫逃逸。
